A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters.

janelia7_blocks-janelia7_biblio_header | block

Nature Chemical Biology. 2018 Feb 26;. doi: 10.1038/s41589-018-0004-9

A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters. Ahrens Lab

Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk H, Hochbaum DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang C, Rhoades JL, Shemesh O, Asano S, Yoon Y, Freifeld L, Saulnier JL, Riegler C, Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, Sabatini BL, Boyden ES

janelia7_blocks-janelia7_biblio_abstract | block

Abstract

We developed a new way to engineer complex proteins toward multidimensional specifications using a simple, yet scalable, directed evolution strategy. By robotically picking mammalian cells that were identified, under a microscope, as expressing proteins that simultaneously exhibit several specific properties, we can screen hundreds of thousands of proteins in a library in just a few hours, evaluating each along multiple performance axes. To demonstrate the power of this approach, we created a genetically encoded fluorescent voltage indicator, simultaneously optimizing its brightness and membrane localization using our microscopy-guided cell-picking strategy. We produced the high-performance opsin-based fluorescent voltage reporter Archon1 and demonstrated its utility by imaging spiking and millivolt-scale subthreshold and synaptic activity in acute mouse brain slices and in larval zebrafish in vivo. We also measured postsynaptic responses downstream of optogenetically controlled neurons in C. elegans.

node:body | entity_field

Main Menu (Mobile)- Block

Main Menu - Block

A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters.

Abstract

Janelia Authors